Cross-reactivity of IgE antibodies to allergens

The cross‐reactivity of IgE antibodies is of interest for various reasons, three of which are discussed. Firstly, from the clinical view, it is important to know the patterns of cross‐reactivity, because they often (but not always) reflect the pattern of clinical sensitivities. We discuss the cross‐reactivities associated with sensitization to pollen and vegetable foods: PR‐10 (Bet v 1‐related), profilin, the cross‐reactive carbohydrate determinant (CCD), the recently described isoflavone reductase, and the (still elusive) mugwort allergen that is associated with celery anaphylaxis; cross‐reactivities between allergens from invertebrates, particularly tropomyosin, paramyosin, and glutathione S‐transferase (GST); and latex‐associated cross‐reactivities. Clustering cross‐reactive allergens may simplify diagnostic procedures and therapeutic regimens. Secondly, IgE cross‐reactivity is of interest for its immunologic basis, particularly in relation to the regulation of allergic sensitization: are IgE antibodies to allergens more often cross‐reactive than IgG antibodies to “normal” antigens? If so, why? For this discussion, it is relevant to compare not only the structural relation between the two allergens in question, but also the relatedness to the human equivalent (if any) and how the latter influences the immune repertoire. Thirdly, prediction of IgE cross‐reactivity is of interest in relation to allergic reactivity to novel foods. Cross‐reactivity is a property defined by individual antibodies to individual allergens. Quantitative information (including relative affinity) is required on cross‐reactivity in the allergic population and with specific allergens (rather than with whole extracts). Such information is still scarce, but with the increasing availability of purified (usually recombinant) allergens, such quantitative information will soon start to accumulate. It is expected that similarity in short stretches of the linear amino‐acid sequence is unlikely to result in relevant cross‐reactivity between two proteins unless there is similarity in the protein fold.     

Position paper of the EAACI: food allergy due to immunological cross‐reactions with common inhalant allergens

In older children, adolescents, and adults, a substantial part of all IgE‐mediated food allergies is caused by cross‐reacting allergenic structures shared by inhalants and foods. IgE stimulated by a cross‐reactive inhalant allergen can result in diverse patterns of allergic reactions to various foods. Local, mild, or severe systemic reactions may occur already after the first consumption of a food containing a cross‐reactive allergen. In clinical practice, clinically relevant sensitizations are elucidated by skin prick testing or by the determination of specific IgE in vitro. Component‐resolved diagnosis may help to reach a diagnosis and may predict the risk of a systemic reaction. Allergy needs to be confirmed in cases of unclear history by oral challenge tests. The therapeutic potential of allergen immunotherapy with inhalant allergens in pollen‐related food allergy is not clear, and more placebo‐controlled studies are needed. As we are facing an increasing incidence of pollen allergies, a shift in sensitization patterns and changes in nutritional habits, and the occurrence of new, so far unknown allergies due to cross‐reactions are expected.     

Auto‐ and cross‐reactivity to thioredoxin allergens in allergic bronchopulmonary aspergillosis

Background: Thioredoxins are cross‐reactive allergens involved in the pathogenesis of atopic eczema and asthma. Cross‐reactivity to human thioredoxin can contribute to the exacerbation of severe atopic diseases. Methods: Human thioredoxin, Asp f28 and Asp f29, two thioredoxins of Aspergillus fumigatus, and thioredoxin of Malassezia sympodialis were cloned and produced as recombinant proteins. Allergenicity and cross‐reactivity to thioredoxins in allergic bronchopulmonary aspergillosis patients were assessed by enzyme‐linked immunosorbent assay (ELISA), inhibition ELISA, immunoblot analysis, proliferation assays and skin tests. Molecular homology modelling was used to identify conserved, surface‐exposed amino acids potentially involved in immunoglobulin E (IgE)‐binding. Results: All thioredoxins, including the human enzyme, bind IgE from patients with allergic bronchopulmonary aspergillosis and induce allergen‐specific proliferation in peripheral blood mononuclear cells and positive skin reactions in thioredoxin‐sensitized patients. Inhibition experiments showed that the thioredoxins are cross‐reactive indicating humoral immune responses based on molecular mimicry. To identify structural surface elements involved in cross‐reactivity, the three‐dimensional structures were modelled based on solved thioredoxin structures. Analysis of the molecular surfaces combined with sequence alignments allowed identification of conserved solvent exposed amino acids distantly located in the linear sequences which cluster to patches of continuous surface areas. The size of the surface areas conserved between human and fungal thioredoxins correlates well with the inhibitory potential of the molecules in inhibition ELISA indicating that the shared amino acids are involved in IgE‐binding. Conclusions: Conserved, solvent exposed residues shared between different thioredoxins cluster to continuous surface regions potentially forming cross‐reactive conformational B‐cell epitopes responsible for IgE‐mediated cross‐reactivity and autoreactivity.     

Primary versus secondary immunoglobulin E sensitization to soy and wheat in the Multi-Centre Allergy Study cohort

Background IgE sensitization to soy and wheat is classified as ‘primary’ when generated by food ingestion and ‘secondary’ when it as a consequence of primary sensitization to cross‐reacting pollen antigens via inhalation. The age‐specific relevance of these categories of sensitization throughout childhood is unknown. Objective To monitor the natural course of IgE sensitization against common food allergens in childhood in relation to sensitization against cross‐reactive airborne allergens. Methods The German Multi‐Centre Allergy Study with follow‐up from birth to age 13 recruited initially 1314 children. IgE antibody levels against cow's milk, hen's egg, soy, wheat, mites, cat and dog dander, birch and grass pollens were tested. Longitudinal data were analysed from the 273 children with sera obtained at age 2, 5, 7 and 10 years of age. Results The point prevalence of sensitization (>1.0 kU/L) to milk and egg allergens progressively decreased from about 4% at 2 years to <1% at 10 years. By contrast, the prevalence of IgE to wheat and soy progressively increased with age, from 2% to 7% (soy) and from 2% to 9% (wheat). At 10 years of age, IgE to grass pollen was detected in 97% and 98% of the children reacting against soy and wheat, respectively; IgE to birch pollen was observed in 86% and 82% of the children reacting against soy and wheat, respectively. Early IgE sensitization to soy or wheat preceded that to grass or birch pollen in only 4% and 8% of participants sensitized to soy and wheat, respectively. Conclusion IgE sensitization to soy and wheat is relatively uncommon and mostly primary in early infancy, more frequent and mostly secondary to pollen sensitization at school age. Clinical Implications Awareness should be raised to avoid unnecessary diet restrictions due to the high frequency of clinically irrelevant, secondary sensitization to soy and wheat in schoolchildren with pollinosis.     

Cow's milk-specific immunoglobulin E levels as predictors of clinical reactivity in the follow-up of the cow's milk allergy infants

Background IgE‐mediated cow's milk proteins (CMPs) allergy shows a tendency to disappear with age. The sooner tolerance is detected, the earlier the substitute diets can be suspended and the quicker family emotional hardship is alleviated. Objective To analyse the specific IgE levels to cow's milk and its proteins, which help to separate tolerant from no tolerant children in the follow‐up of infants with allergy to cow's milk. Patients and methods Sixty‐six infants diagnosed with IgE‐mediated allergy to CMPs were included in this prospective follow‐up study. Periodic reassessments were carried out every 6 months until they were 2‐years old and then, annually, until tolerance arose or until the last reassessment in which tolerance had not been achieved. Non‐tolerant infants were followed, at least, for a period of 3 years. In each visit, the same skin tests and determination of specific IgE (CAP System FEIA) for milk and its proteins were carried out. The open challenge test was repeated unless a clear transgression to milk, which came to be positive, had taken place within the previous 3 months in each of the follow‐up visits. Specific IgE levels to milk and its proteins, in different moments of the follow‐up were analysed by means of the receiver‐operating characteristic curve to predict clinical reactivity. Results Throughout the follow‐up 45 (68%) infants became tolerant. The follow‐up mean for tolerant infants was 21.2 months whereas for non‐tolerant infants it was 58 months. The specific IgE levels which were predictors of the clinical reactivity (positive predictive value (PPV)90%), grew as the age of the infants increased: 1.5, 6 and 14 kU_A/L for milk in the age range 13–18 and 19–24 months and in the third year, respectively. Specific IgE levels to casein: 0.6, 3 and 5 kU_A/L, respectively, predicted clinical reactivity (PPV90%) in the different analysed moments of the follow‐up. The cut‐off points: 2.7, 9 and 24 kU_A/L for milk and 2, 4.2 and 9 kU_A/L for casein, respectively, predicted clinical reactivity with an accuracy 95% corresponding to a specificity of 90%. Conclusions Monitorization of specific IgE concentration for milk and casein by means of the CAP system in allergic children to CMPs allows us to predict, to a high degree of probability, clinical reactivity. Age factor must be taken into account to evaluate the specific IgE levels which are predictors of tolerance or clinical reactivity.     

Serum immunoglobulin A concentration in infancy,but not human milk immunoglobulin A,is associated with subsequent atopic manifestations in children and adolescents: a 20‐year prospective follow‐up study

Background Serum and secretory IgA concentrations have been suggested to be inversely associated with allergic symptoms in children. Furthermore, low maternal milk IgA concentration has been suggested to be associated with the development of cow's milk allergy. Objective Our aim was to explore whether the serum IgA concentrations in infancy and the IgA concentration of maternal milk predict atopic manifestations in childhood and up to age 20 years. Methods A cohort of 200 unselected full‐term newborns was prospectively followed up from birth to age 20 years with measurement of serum total IgA at ages 2 and 6 months. The mothers were encouraged to maintain exclusive breastfeeding for as long as possible. Total IgA concentration of maternal milk was measured at birth (colostrum, n=169) and at 2 (n=167) and 6 (n=119) months of lactation. The children were re‐assessed at ages 5, 11 and 20 years for the occurrence of allergic symptoms, with skin prick testing and measurement of serum IgE. Results Children and adolescents with respiratory allergic symptoms and sensitization had a higher serum IgA concentration at age 2 months than the non‐atopic subjects. Colostrum and breast milk IgA concentrations were not associated with the development of allergic symptoms in the recipient infant. However, maternal milk IgA concentration at 6 months of lactation was inversely associated with elevated serum total IgE and positive skin prick test to tree pollen in the offspring at age 20 years. Conclusions and Clinical Relevance Increased serum IgA concentration at age 2 months is associated with the development of subsequent allergic symptoms and sensitization in childhood and adolescence. Maternal milk IgA concentrations are not associated with subsequent allergic symptoms in the recipient infant. The present study provides novel information on the role of IgA in the development of respiratory allergy and sensitization. Cite this as: M. Pesonen, M. J. T. Kallio, M. A. Siimes, E. Savilahti and A. Ranki, Clinical & Experimental Allergy, 2011 (41) 688–696.     

Characterization of peptidic and carbohydrate cross-reactive determinants in pollen polysensitization

Background Cross‐reactivity may be due to protein sequence or domain homologies and/or the existence of cross‐reactive carbohydrate determinants (CCDs). The clinical relevance of peptidic cross‐reactivities is well known, whereas that of CCDs is still a question of debate. The aim of this study is to characterize the IgE specificity of various patients suffering from pollen polysensitization to identify both peptidic and carbohydrate cross‐reactive determinants. Material and methods Rapeseed, grass and Arabidopsis proteins were separated by isoelectric focusing, followed by SDS‐PAGE, and transferred to a nitrocellulose sheet. The sheets were incubated either with an individual serum from a birch+grass‐sensitive patient, followed by anti‐human IgE, or with labelled Concanavalin A (ConA). Binding inhibition was tested by incubation of the sera with a mixture of sugar residues. Results The results showed two different patterns of cross‐reacting sera: a pattern that implies few proteins, not always glycosylated and known as allergens, and a pattern that implies numerous proteins with molecular masses over 30 kDa. This second pattern was very close to the ConA ‐binding pattern. The IgE binding was abolished by pre‐incubation with sugar residues only in the case of the second pattern. Discussion This study shows that multiple pollen sensitizations could result from multiple sensitizations to specific proteins or from a cross‐sensitization to a wide range of glycoproteins. Two‐D blots allow to characterize a cross‐sensitization due to carbohydrate determinants, and thus to improve the diagnosis of allergy and its medical treatment.     

Allergy to goat and sheep milk without allergy to cow's milk

BACKGROUND: Cow's milk (CM) allergy is the most frequent cause of food allergy in infants. Most children who are allergic to CM are also sensitized to whey proteins and/or to the casein fraction and many of them cannot tolerate goat's or sheep's milk (GSM) either. Conversely, the GSM allergies that are not associated with allergic cross-reactivity to CM are rare. METHODS: Twenty-eight children who had severe allergic reactions, including anaphylaxis, after consumption of GSM products but tolerated CM products were recruited in a retrospective study. Whole casein and whey proteins were fractionated from CM and GSM. beta-Lactoglobulin and the different caseins were isolated, purified and used to perform enzyme allergosorbent tests (EAST) and EAST inhibition studies with the sera of the allergic children. RESULTS: Clinical observations, skin prick testing and immunoglobulin (Ig)E-binding studies confirmed the diagnosis of GSM allergy without associated CM allergy. EAST determinations demonstrated that GSM allergy involves the casein fraction and not whey proteins. Cow's milk caseins were not at all or poorly recognized by the patient's IgE, while alphaS(1)-, alphaS(2)- and beta-caseins from GSM were recognized with a high specificity and affinity. In all cases, increasing concentrations of CM caseins failed to inhibit the binding of patient's IgE to sheep or goat milk caseins, whereas this binding was completely inhibited by GSM caseins. CONCLUSIONS: The characteristics of GSM allergy differ from those of the CM allergy because it affects older children and appears later. CM products do not elicit any clinical manifestation in GSM allergic patients, whereas CM allergic patients, usually cross-react to GSM. In all the GSM allergic children, the IgE antibodies recognized the caseins but not the whey proteins. Moreover, IgE specificity and affinity was high to GSM and lower to CM caseins despite their marked sequence homology. Doctors and allergic individuals should be aware that GSM allergy requires a strict avoidance of GSM and milk-derived products because reactions could be severe after ingestion of minimal doses of the offending food.     

A new framework for the interpretation of IgE sensitization tests

IgE sensitization tests, such as skin prick testing and serum‐specific IgE, have been used to diagnose IgE‐mediated clinical allergy for many years. Their prime drawback is that they detect sensitization which is only loosely related to clinical allergy. Many patients therefore require provocation tests to make a definitive diagnosis; these are often expensive and potentially associated with severe reactions. The likelihood of clinical allergy can be semi‐quantified from an IgE sensitization test results. This relationship varies though according to the patients’ age, ethnicity, nature of the putative allergic reaction and coexisting clinical diseases such as eczema. The likelihood of clinical allergy can be more precisely estimated from an IgE sensitization test result, by taking into account the patient's presenting features (pretest probability). The presence of each of these patient‐specific factors may mean that a patient is more or less likely to have clinical allergy with a given test result (post‐test probability). We present two approaches to include pretest probabilities in the interpretation of results. These approaches are currently limited by a lack of data to allow us to derive pretest probabilities for diverse setting, regions and allergens. Also, cofactors, such as exercise, may be necessary for exposure to an allergen to result in an allergic reaction in specific IgE‐positive patients. The diagnosis of IgE‐mediated allergy is now being aided by the introduction of allergen component testing which may identify clinically relevant sensitization. Other approaches are in development with basophil activation testing being closest to clinical application.     

Pru p 3, a marker allergen for lipid transfer protein sensitization also in Central Europe

In the Mediterranean area, lipid transfer proteins (LTPs) are important causes of plant‐food allergies often associated with severe allergic reactions. There, peach LTP (Pru p 3) seems to be the primary sensitizer, whereas in Central Europe, little is known about the importance of LTP sensitization. In this region, allergen extract‐based diagnosis is often complicated by co‐sensitization to Bet v 1, the major birch pollen allergen, its cross‐reactive food allergens, and profilins. We investigated the role of LTP sensitization in Central European patients displaying strong allergic reactions to plant‐derived food. Analysis of IgE reactivity revealed that ten of thirteen patients were sensitized to Pru p 3, nine to Bet v 1, and two to profilin. Our results showed that LTP sensitization represents a risk factor for severe allergic symptoms in Central Europe. Furthermore, the strong IgE reactivity detected in immunoblots of plant‐food extracts indicated that Pru p 3 can be used as a marker allergen for LTP sensitization also in Central European patients.     

Animal-derived lipocalin allergens exhibit immunoglobulin E cross-reactivity

Background Although knowledge of the IgE cross‐reactivity between allergens is important for understanding the mechanisms of allergy, the regulation of the allergic immune response and the development of efficient modes of allergen immunotherapy, the cross‐reactivity of animal allergens is poorly known. Objective The aim of this study was to characterize IgE cross‐reactivities between lipocalin proteins, including five animal‐derived lipocalin allergens and one human endogenous lipocalin, tear lipocalin (TL). Methods The recombinant proteins were validated by chromatography and mass spectrometry. The IgE‐binding capacity of the allergens was confirmed by IgE. immunoblotting and IgE immunoblot inhibition. IgE ELISA was performed with sera from 42 atopic patients and 21 control subjects. The IgE cross‐reactivities between the lipocalin proteins were determined by ELISA inhibition. Results ELISA inhibition revealed IgE cross‐reactivities between Can f 1 and human TL, between Can f 1 and Can f 2, and between Equ c 1 and Mus m 1. Low levels of IgE to human TL were found in the sera of seven dog‐allergic patients of whom six were IgE‐positive for Can f 1. Conclusion Several lipocalins exhibited IgE cross‐reactivity, probably due to the sequential identity of the proteins and also due to similarities in their three‐dimensional structures. The clinical significance of the findings needs to be elucidated. Low‐level IgE cross‐reactivity can play a role in regulating immune response to lipocalin allergens.     

The cat lipocalin Fel d 7 and its cross‐reactivity with the dog lipocalin Can f 1

We investigated the prevalence of sensitization to the cat lipocalin Fel d 7 among 140 cat‐sensitized Swedish patients and elucidated its allergenic activity and cross‐reactivity with the dog lipocalin Can f 1. Sixty‐five of 140 patients had IgE to rFel d 7 whereof 60 also had IgE to rCan f 1. A moderate correlation between IgE levels to rFel d 7 and rCan f 1 was found. rFel d 7 activated basophils in vitro and inhibited IgE binding to rCan f 1 in 4 of 13 patients, whereas rCan f 1 inhibited IgE binding to rFel d 7 in 7 of 13 patients. Fel d 7 and Can f 1 showed high similarities in protein structure and epitopes in common were found using cross‐reactive antisera. Fel d 7 is a common allergen in a Swedish cat‐sensitized population that cross‐reacts with Can f 1, and may contribute to symptoms in cat‐ but also in dog‐allergic patients.     

The allergen profile of beech and oak pollen

Background Beech and oak pollen are potential allergen sources with a world‐wide distribution. Objective We aimed to characterize the allergen profile of beech and oak pollen and to study cross‐reactivities with birch and grass pollen allergens. Methods Sera from tree pollen‐allergic patients with evidence for beech and oak pollen sensitization from Basel, Switzerland, (n=23) and sera from birch pollen‐allergic patients from Vienna, Austria, (n=26) were compared in immunoblot experiments for IgE reactivity to birch (Betula pendula syn. verrucosa), beech (Fagus sylvatica) and oak (Quercus alba) pollen allergens. Subsequently, beech and oak pollen allergens were characterized by IgE inhibition experiments with purified recombinant and natural allergens and with allergen‐specific antibody probes. Birch‐, beech‐ and oak pollen‐specific IgE levels were determined by ELISA. Results Beech and oak pollen contain allergens that cross‐react with the birch pollen allergens Bet v 1, Bet v 2 and Bet v 4 and with the berberine bridge enzyme‐like allergen Phl p 4 from timothy grass pollen. Sera from Swiss and Austrian patients exhibited similar IgE reactivity profiles to birch, beech and oak pollen extracts. IgE levels to beech and oak pollen allergens were lower than those to birch pollen allergens. Conclusion IgE reactivity to beech pollen is mainly due to cross‐reactivity with birch pollen allergens, and a Phl p 4‐like molecule represented another predominant IgE‐reactive structure in oak pollen. The characterization of beech and oak pollen allergens and their cross‐reactivity is important for the diagnosis and treatment of beech and oak pollen allergy.     

The impact of caesarean delivery and type of feeding on cow’s milk allergy in infants and subsequent development of allergic march in childhood

Background: The incidence of IgE‐mediated cow’s milk allergy (CMA) has increased over the last few years. There are several genetic and environmental risk factors that may be related to this allergy and the subsequent allergic march (AM). Methods: A prospective, cohort study was conducted in patients recruited into the study between 1998 and 2002. Information on clinical variables and complementary tests, perinatal and obstetric factors and the type of hydrolysed formula used was recorded. A cross sectional study on the prevalence of allergic diseases in this cohort was performed in 2004. Results: We compared IgE‐mediated CMA patients with non‐IgE‐mediated CMA patients and found that IgE‐mediated CMA is associated with caesarean delivery (OR = 2.14 95% CI: 1.02–4.49), duration of breast feeding (>2 months, OR = 4.14; 95% CI: 2.17–7.89) and the use of supplementary artificial formula whilst breast feeding (OR = 2.86; 95% CI: 1.33–6.13). The factors associated with AM in IgE‐mediated CMA patients were caesarean delivery (OR = 0.42; 95% CI: 0.19–0.92) and the use of more extensively hydrolysed high grade hydrolysates (+EH/HGH) (OR = 0.44; 95% CI: 0.20–0.98), both as protective factors. Conclusions: Caesarean delivery is demonstrated as being a risk factor for IgE‐mediated CMA, but it does not increase the risk of AM in these infants. The use of +EH/HGH appears to protect IgE‐mediated CMA patients from eventually developing AM.     

Impact of cross‐reactive carbohydrate determinants on wood dust sensitization

Background Occupational wood dust exposure can induce allergy and may be one cause of respiratory health problems among woodworkers. Objective The objective was to determine the prevalence and quantitative level of specific immunoglobulin E (sIgE) to beech and pine wood in exposed workers. Wood sensitization was specified with regard to cross‐reactivity and was correlated to the reported symptoms. Methods Danish workers (n=701) were investigated for sIgE to beech and pine. Wood samples from workplaces were analysed and coupled to ImmunoCAPs. Workers sensitized to wood were tested for cross‐reactive carbohydrate determinants (CCDs) and environmental allergens. IgE binding was specified for glycogenic vs. proteinogenic epitopes by inhibition tests. Results The prevalence of wood sensitization among all workers was 3.7%. There was no association between sensitization prevalence or sIgE concentrations and self‐reported allergic symptoms. Beech‐ and pine‐sensitized workers showed a high prevalence of CCD sensitization (73%). However, workers with a single sensitization to wood had no sIgE to CCDs. Specifying IgE epitopes demonstrated that sera of workers reporting allergic symptoms recognized proteinogenic IgE‐epitopes on wood allergens, whereas workers without allergic symptoms had primarily sIgE‐epitopes to glycogenic structures. Although 96% of the wood‐sensitized workers were atopic, no significant correlation was found between wood sensitization and sIgE to beech and birch pollen, but an association was found between sIgE against CCDs and pine pollen. Conclusion Sensitization prevalence to beech and pine wood measured by tailored ImmunoCAPs was not correlated to allergic symptoms. We recommend the application of CCD tools to assess the relevance of individual wood sensitization. Cite this as: S. Kespohl, V. Schlünssen, G. Jacobsen, I. Schaumburg, S. Maryska, U. Meurer, T. Brüning, T. Sigsgaard and M. Raulf‐Heimsoth, Clinical & Experimental Allergy, 2010 (40) 1099–1106.     

Component‐resolved diagnostics to direct in venom immunotherapy: Important steps towards precision medicine

Stings of Hymenoptera can induce IgE‐mediated systemic and even fatal allergic reactions. Venom‐specific immunotherapy (VIT) is the only disease‐modifying and curative treatment of venom allergy. However, choosing the correct venom for VIT represents a necessary prerequisite for efficient protection against further anaphylactic sting reactions after VIT. In the past, therapeutic decisions based on the measurement of specific IgE (sIgE) levels to whole venom extracts were not always straightforward, especially when the patient was not able to identify the culprit insect. In the last years, the increasing knowledge about the molecular structure and relevance of important venom allergens and their availability as recombinant allergens, devoid of cross‐reactive carbohydrate determinants, resulted in the development of an advanced component‐resolved diagnostics (CRD) approach in venom allergy. Already to date, CRD has increased the sensitivity of sIgE detection and enabled the discrimination between primary sensitization and cross‐reactivity, particularly in patients with sensitization to both honeybee and vespid venom. Hence, CRD in many patients improves the selection of the appropriate immunotherapeutic intervention. Moreover, the detailed knowledge about sensitization profiles on a molecular level might open new options to identify patients who are at increased risk of side‐effects or not to respond to immunotherapy. Therefore, increasing potential of CRD becomes evident, to direct therapeutic decisions in a personalized and patient‐tailored manner. Reviewed here are the state of the art options, recent developments and future perspectives of CRD of Hymenoptera venom allergy.     

Immunochemical characterization of Glycine max L. Merr. var Raiden, as a possible hypoallergenic substitute for cow's milk-allergic patients

Background Cows' milk allergy (CMA) is the most common cause of food allergy in infancy. The only proven treatment is the complete elimination of cows' milk proteins (CMPs) from the diet by means of hypoallergenic formulas. Soybean‐based formulae are widely used although intolerance to soy has been reported to occur in 15–40% of infants with CMA. Objective The aim of this work was to analyse the in vitro reactivity of the soybean cultivar Raiden, which naturally lacks glycinin A₄A₅B₃, to evaluate whether this genotype could be a safe CMP substitute for CMA patients. Methods The reactivity of conventional soybean (CS) and Raiden soybean (RS) genotypes and also recombinant glycinin A₄A₅B₃ and αβ‐conglycinin with casein‐specific monoclonal antibodies and CMP‐specific polyclonal serum was evaluated by immunoblotting and ELISA. A sequential competitive ELISA with the polyclonal antiserum and different soluble inhibitors was performed. In addition, an indirect ELISA with sera of atopic children with CMA was carried out to analyse the IgE‐binding capacity of the different soybean components. Results We have shown that CS contains four components that cross‐react with CMP, while RS has only one. The remaining cross‐reactive component in RS was identified as α‐subunit β‐conglycinin. By means of inhibitory ELISA, we demonstrated that CS, RS and the α‐subunit β‐conglycinin extracts inhibited the binding of CMP‐specific antibodies to the CMP‐coated solid phase. Finally, we showed that CS, RS and the recombinant proteins were recognized by human CMP‐specific IgE antibodies. Conclusion This work shows that although Raiden has fewer cross‐reactive components than conventional soybean, it still has a residual cross‐reactive component: the α‐subunit β‐conglycinin. This reactivity might make this genotype unsuitable to treat CMA and also explains adverse reactions to soybean in CMA infants.     

Maize food allergy: a double‐blind placebo‐controlled study

Background Maize allergy is not very common especially in Europe. The number of studies that address IgE mediated maize allergy is all too few. Objective Evaluate subjects with a history of maize allergy by double‐blind, placebo‐controlled food challenge; identify the spectrum of symptoms manifested during challenge; determine the lowest provocation dose (PD) during challenge; determine the performance characteristics of maize skin prick test and specific IgE. Methods Twenty‐seven patients with a history of maize allergy were enrolled to be evaluated by skin test, specific IgE and double‐blind placebo‐controlled maize challenge. Results Forty‐eight percent of the patients were challenge positive. PD range was 0.1–25 g. Fifty‐four percent of the maize allergic subjects had a PD that was 2.5 g; two subjects reacted to 100 mg of maize. Comparison of maize specific IgE levels and skin test results to the challenge results revealed the following (specific IgE level/skin testing): sensitivity 1.00/0.846, specificity 0.077/0.384, positive predictive value 0.520/0.579, and negative predictive value 1.00/0.714. Conclusion Maize is a cause of IgE‐mediated allergic reactions to foods in adults and children. Nearly half of the subjects recruited were confirmed by challenge to be allergic to maize. Twenty‐three percent of the positive challenge patients manifested symptoms that involved two organ systems, thus fulfilling the criteria for maize induced anaphylaxis. Maize is allergenic and can pose a risk for symptomatic food allergy at a dose of 100 mg.     

Allergenicity of major cow's milk and peanut proteins determined by IgE and IgG immunoblotting

BACKGROUND: Specific IgG antibodies are frequently observed in food-allergic patients. However, the allergen-fraction specificity of IgG antibodies in relation to IgE antibodies is not well defined. Our aim was to determine the IgE and IgG antibody profile to major cow's milk and peanut-antigen fractions in food-allergic patients and tolerant individuals. METHODS: Sera were collected from 10 patients allergic to cow's milk and 10 patients allergic to peanuts, as well as from 20 control subjects. Cow's milk and peanut proteins were migrated on SDS-PAGE and immunoblotted for IgE, IgG, and IgG4 antibodies. Food-specific IgE concentrations were measured by CAP System FEIA, and IgG and IgG4 concentrations by ELISA. RESULTS: In food-allergic children, similar fraction-specific IgE, IgG, and IgG4 antibody-binding profiles to the major cow's milk or peanut antigens were found. In nonallergics, the presence of fraction-specific IgG antibodies was mostly dependent on regular ingestion of the food. The presence of specific antibody on immunoblots correlated with their quantitative measurement. The mean value for specific IgE in cow's milk-allergic patients was 450 +/- 1,326 IU/ml, and 337 +/- 423 IU/ml in peanut-allergic patients. Specific IgG antibody values in milk-allergic patients were not different (median OD 1.5, range 0.3-2.3) from controls (median OD 1, range 0.2-1.8). However, in peanut-allergic patients, IgG concentrations were significantly higher than in controls (OD 1.2 [0.5-1.3] vs 0.5 [0.3-0.7]; P< 0.01). CONCLUSIONS: Similar fraction-specific IgE and IgG antibody profiles in allergic individuals suggest a common switching trigger involving both isotypes. Intrinsic allergenicity might explain identical IgG antibody fraction specificity in nonallergics and in allergics. The presence of IgG antibodies in nonallergics was related to regular ingestion of the food.     

Diagnostic significance of in vitro T-cell proliferative responses to house-dust mite Der p 1 in children with dust-mite allerev

The study aimed to investigate the possible diagnostic significance of T-cell proliferative responses to purified Der p 1 antigen in allergic children with and without allergic sensitization to house-dust-mite (HDM) allergens. T-cell reactivity to purified Der p 1antigen was analyzed in 24 children with allergic sensitization to HDM demonstrated by RAST and/or positive skin prick tests. Twelve healthy young adults and 11 children with allergic sensitizations other than HDM served as controls. Of 25 HDM-allergic patients, 16 displayed strong T-cell reactivity to Der p 1 (stimulation indices >3): nine patients showed no T-cell proliferation despite the presence of specific IgE, and five showed no responses despite positive skin prick test. In two patients, a weak T-cell response to Der p 1 could be demonstrated in the absence of specific IgE and negative skin test result. The two affected subjects did not show evidence of mite allergy. No T-cell responses were observed in adult controls (stimulation indices<3). We conclude that the assessment of T-cell reactivity to Der p 1 is of little value for the diagnosis of HDM allergy. The importance of T-cell proliferative responses for the study of the pathogenesis of HDM allergy remains unchallenged.